Anti-TORC1/CRTC1 Antibody Picoband™

Figure 1. Western blot analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: human U20S whole cell lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TORC1/CRTC1 antigen affinity purified polyclonal antibody (Catalog # A01951-3) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TORC1/CRTC1 at approximately 78KD. The expected band size for TORC1/CRTC1 is at 78KD.

Figure 2. IHC analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
TORC1/CRTC1 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TORC1/CRTC1 Antibody (A01951-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
TORC1/CRTC1 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TORC1/CRTC1 Antibody (A01951-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
TORC1/CRTC1 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TORC1/CRTC1 Antibody (A01951-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 5. IHC analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
TORC1/CRTC1 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TORC1/CRTC1 Antibody (A01951-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 6. IF analysis of TORC1/CRTC1 using anti-TORC1/CRTC1 antibody (A01951-3).
TORC1/CRTC1 was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-TORC1/CRTC1 Antibody (A01951-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 7. Flow Cytometry analysis of A431 cells using anti-TORC1/CRTC1 antibody (A01951-3).
Overlay histogram showing A431 cells stained with A01951-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TORC1/CRTC1 Antibody (A01951-3, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 8. Flow Cytometry analysis of RH35 cells using anti-TORC1/CRTC1 antibody (A01951-3).
Overlay histogram showing RH35 cells stained with A01951-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TORC1/CRTC1 Antibody (A01951-3, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.